Department of Chemistry, Gwangju Institute of Science and Technology, 123 Cheomdangwagi-ro, Buk-gu, Gwangju 61005, Korea.
Sensors (Basel). 2019 Jun 18;19(12):2728. doi: 10.3390/s19122728.
Adenosine deaminase (ADA), which catalyzes the irreversible deamination of adenosine to inosine, is related to various human diseases such as tuberculous peritonitis and leukemia. Therefore, the method used to detect ADA activity and screen the effectiveness of various inhibitor candidates has important implications for the diagnosis treatment for various human diseases. A simple and rapid assay method for ADA, based on the enzymatic formation of a luminescent lanthanide complex, is proposed in this study. Inosine, an enzymatic product of ADA with stronger sensitization efficiency for Tb than adenosine, produced a strong luminescence by forming an inosine-Tb complex, and it enabled the direct monitoring of ADA activity in real-time. By introducing only Tb to adenosine and ADA in the buffer, the enhancement of luminescence enabled the detection of a low concentration of ADA (detection limit 1.6 U/L). Moreover, this method could accurately determine the inhibition efficiency (IC) of the known ADA inhibitor, erhythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), and the inhibition of ADA could be confirmed by the naked eye. Considering its simplicity, this assay could be extended to the high-throughput screening of various ADA inhibitor candidates.
腺苷脱氨酶（ADA）可催化腺苷不可逆脱氨为肌苷，与结核性腹膜炎和白血病等多种人类疾病有关。因此，用于检测 ADA 活性和筛选各种抑制剂候选物的有效性的方法对于各种人类疾病的诊断和治疗具有重要意义。本研究提出了一种基于酶促形成发光镧系配合物的 ADA 简单快速检测方法。ADA 的酶产物肌苷比腺苷对 Tb 的敏化效率更强，通过形成肌苷-Tb 配合物产生强烈的发光，从而能够实时直接监测 ADA 活性。通过仅在缓冲液中向腺苷和 ADA 中引入 Tb，发光的增强能够检测到低浓度的 ADA（检测限 1.6 U/L）。此外，该方法可以准确地确定已知 ADA 抑制剂（erythro-9-（2-羟基-3-壬基）腺嘌呤（EHNA）的抑制效率（IC），并且可以通过肉眼确认 ADA 的抑制作用。考虑到其简单性，该测定法可扩展到各种 ADA 抑制剂候选物的高通量筛选。
